Project description:Genome wide DNA methylation profiling of normal and squamous carcinoma of the cervix. The Illumina Infinium 27k Human DNA methylation Beadchip v1.2 was used to obtain DNA methylation profiles across approximately 27,000 CpGs in frozen cervical samples. Samples included 4 normal ectocervices (Ecto 1 to 4) and 5 squamous cell carcinoma of the cervix (SCC 1 to 5). In order to identify epigenetic biomarkers for early cervix cancer diagnosis, we performed a methylation screening using Illumina Infinium 27k Human DNA methylation Beadchip v1.2 of stem cell marker promoters during cervical carcinogenesis and demonstrated a strong hypermethylation of Undifferentiated cell Transcription Factor 1 (UTF1) promoter in squamous cell carcinoma (SCC) in comparison with normal ectocervix. Direct bisulfite pyrosequencing of DNA isolated from liquid-based cytological samples showed that UTF1 promoter methylation increases with lesion severity and is associated with higher expression. By RT-PCR, Western Blot and immunofluorescence, we demonstrated that UTF1 mRNA and protein are expressed in epithelial cancer cell lines, even in the absence of its two main described regulators Oct4A and Sox2. Methyl-Specific PCR experiments revealed that the inhibition of DNA methyltransferase by 5-aza-2’-deoxycytidine is associated with decreased UTF1 gene methylation and expression. These findings provide evidence that UTF1 promoter methylation profile might be a useful biomarker for cervix cancer diagnosis and raise the questions of its role during epithelial carcinogenesis and of the mechanisms involved in the regulation of its expression. Bisulfite converted DNA from the 9 samples were hybridised to the Illumina Infinium 27k Human Methylation Beadchip v1.2
Project description:Gene methylation profiling of immortalized human mesenchymal stem cells comparing HPV E6/E7-transfected MSCs cells with human telomerase reverse transcriptase (hTERT)- and HPV E6/E7-transfected MSCs. hTERT may increase gene methylation in MSCs. Goal was to determine the effects of different transfected genes on global gene methylation in MSCs.
Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
Project description:In this study, we characterize the fusion protein produced by the EPC1-PHF1 translocation in Low Grade Endometrial Stromal Sarcoma (LG-ESS) and Ossifying FibroMyxoid Tumors (OFMT). We express the fusion protein and necessary controls in K562 Cells. The fusion protein assembles a mega-complex harboring both NuA4/TIP60 and PRC2 subunits and enzymatic activities and leads to mislocalization of chromatin marks in the genome, linked to aberrant gene expression.
Project description:Gene methylation profiling of immortalized human mesenchymal stem cells comparing HPV E6/E7-transfected MSCs cells with human telomerase reverse transcriptase (hTERT)- and HPV E6/E7-transfected MSCs. hTERT may increase gene methylation in MSCs. Goal was to determine the effects of different transfected genes on global gene methylation in MSCs. Two-condition experiment, KP MSCs vs. 3A6 MSCs.
Project description:In this study, we characterize the fusion protein produced by the EPC1-PHF1 translocation in Low Grade Endometrial Stromal Sarcoma (LG-ESS) and Ossifying FibroMyxoid Tumors (OFMT). We express the fusion protein and necessary controls in K562 Cells. The fusion protein assembles a mega-complex harboring both NuA4/TIP60 and PRC2 subunits and enzymatic activities and leads to mislocalization of chromatin marks in the genome, linked to aberrant gene expression.
Project description:In this study, we characterize the fusion protein produced by the EPC1-PHF1 translocation in Low Grade Endometrial Stromal Sarcoma (LG-ESS) and Ossifying FibroMyxoid Tumors (OFMT). We express the fusion protein and necessary controls in K562 Cells. The fusion protein assembles a mega-complex harboring both NuA4/TIP60 and PRC2 subunits and enzymatic activities and leads to mislocalization of chromatin marks in the genome, linked to aberrant gene expression.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.